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Analysis of CA9 (Human) by ELISA (CAT#: STEM-MB-0710-LGZ)

Introduction

Carbonic anhydrase (CA) is an ancient family of zinc metalloenzymes present in almost all organisms. All CAs can be grouped into 3 broad classes (α, β, and γ), which have evolved independently and also vary widely in sequence and overall folding.
CA9, a member of the alpha class, is a plasma membrane protein with a catalytic domain in the extracellular space. Its expression is restricted to very few healthy tissues (mainly the gastrointestinal tract). CA9 expression is strongly induced by hypoxia and can be downregulated by wild-type von Hippel–Lindau (VHL) tumor suppressor protein. CA9 expression is elevated in many types of tumors, especially solid hypoxic tumors that do not respond well to conventional radiation and/or chemotherapy; CA9 is considered a marker of tumor hypoxia and a promising target for cancer therapeutic intervention.




Principle

Enzyme-linked immunosorbent assay (ELISA) is an enzyme-labeled solid phase immunoassay technique. Its basic principle is to bind the antigen (or antibody) to the solid phase carrier, and the antigen (or antibody) and a certain enzyme link to enzyme labeled antigen (or antibody). During detection, the sample to be tested and the enzymic antigen (or antibody) react with the antigen (or antibody) on the solid phase carrier according to certain procedures, and then remove the unreacted part by washing method. After adding the substrate, the substrate is catalyzed by the enzyme on the solid phase carrier to produce colored substances. Through qualitative or quantitative detection of the amount of colored products, the content of the substance to be measured in the sample can be determined.

Applications

Kidney Injury, Oncology & Cancer, Toxicology

Procedure

1. Add standards or samples to each well and incubate.
2. Pour off the liquid in the well, biotinylated antibody working solution and incubate.
3. Add enzyme conjugate working solution and incubate.
4. Add substrate TMB and incubate.
5. Add stop solution and measure OD value.
6. Calculation of results.

Materials

• Sample Type: Serum, plasma or supernatant
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